Fig. 4.

Nck1 is involved in Schwann cell precursor migration by PDGF whose receptor associates with the cognate receptor through Nck1's SH2 domain. (A, B) Schwann cell precursors were transfected with Nck1 or control luciferase siRNA. Cell lysates were used for an immunoblotting using the respective antibodies against Nck1 or the control actin or Dock8. The scanned bands were densitometrically analyzed. Data were evaluated using Student's t-test (**, p<0.01; n=3). (C, D) The reaggregates of Schwann cell precursors, which were transfected with Nck1 or control siRNA, were placed on dishes and were allowed to migrate in the presence or absence of 20 ng/ml PDGF. Data were evaluated using one-way ANOVA (**, p<0.01; n=4). Scale bars indicate 100 µm. (E) The lysates of rat neonatal sciatic nerve tissues were affinity-precipitated with an immobilized GST-tagged Nck1 (the wild type or the mutant harboring SH2 domain-defective R308K) protein. The precipitates were also immunoblotted with an anti-GST antibody to detect GST-Nck1. The immunoblot for the total PDGF receptor (designated as PDGFR in the panel) was also shown.